1,1,2,2,3,3,4,4,4-nonafluoro-N-(2-hydroxyethyl)-N-methylbutane-1-sulphonamide

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study was conducted in compliance with OECD GLP regulations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 422 (Combined Repeat Dose Toxicity with the Reproduction/Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

T-7599.7

IUPAC Name:

T-7599.7

Test material form:

other: waxy solid

Details on test material:

- Name of test material (as cited in study report): T-7599.7
- Molecular formula (if other than submission substance):
- Molecular weight (if other than submission substance):
- Smiles notation (if other than submission substance):
- InChl (if other than submission substance):
- Structural formula attached as image file (if other than submission substance): see Fig.
- Substance type: Mono constituent substance
- Physical state: Waxy solid
- Analytical purity: 95.55
- Impurities (identity and concentrations):
- Composition of test material, percentage of components:
- Isomers composition:
- Purity test date: 30 January 2002
- Lot/batch No.: Lot 6
- Expiration date of the lot/batch:
- Radiochemical purity (if radiolabelling):
- Specific activity (if radiolabelling):
- Locations of the label (if radiolabelling):
- Expiration date of radiochemical substance (if radiolabelling):
- Stability under test conditions:
- Storage condition of test material: Stored at room temperature
- Other:

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleight, North Carolina
- Age at study initiation: Males: 71 days, Females: 65 days
- Weight at study initiation: Males: 282-333 g, Females: 185-226 g
- Fasting period before study: None
- Housing: F0 generation rats were individually housed in stainless steel wire-bottomed cages except during cohabitation period and postpartum periods. During cohabitation, each pair of male and female rats was housed in the male rat's cage. No later than DG 20, F0 generation female rats were individually housed in nesting boxes.
- Diet (e.g. ad libitum): Certified Rodent Diet #5002 (PMI Nutrition International, Inc., St. Louis, Missouri)
- Water (e.g. ad libitum): Reverse osmosis water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 18 February 2002 To: 25 March 2002

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): Test article solubility
- Concentration in vehicle: 0, 1, 5, 25 mg test article/mL vehicle
- Amount of vehicle (if gavage): Dose volume was 10 mL/kg body weight
- Lot/batch no. (if required): Lot120K0252

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration and homogeneity of the prepared formulations. Quadruplicate samples (2 mL) will be taken from the top, middle and bottom of each concentration on the first day of preparation. Two of those samples will be shipped for analysis.

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 Days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as Day 0 of pregnancy
- After 7 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- After successful mating each pregnant female was caged (how): individually in nesting boxes.

Duration of treatment / exposure:

F0 Generation Rats: Male rats were administered the test substance and/or the vehicle once daily beginning 14 days before cohabitation (maximum 14 days) and continuing until sacrifice, after completion of the cohabitation period, after a minimum of 28 days of dosage. Female rats were administered the test substance and/or the vehicle once daily beginning 14 days before cohabitation (maximum 14 days) and continuting until DL 5.

Frequency of treatment:

Daily

Duration of test:

Each animal recieved a minimum of 28 doses.
IN-LIFE DATES: From: 18 February 2002 To: 25 March 2002

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 10, 50, 250 mg/kg
Basis:
actual ingested

No. of animals per sex per dose:

15

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on previous studies with the test substance.
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: None

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily for viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Hourly on the first day of dosage and daily therafter an hour after dosing. Female rats were evaluated for adverse clinical signs observed during parturition, duration of gestation (DG 0 to the day the first pup was observed), litter sizes (all pups delivered) and pup viability at birth. The pups in each litter were counted once daily. Clinical observations were recorded once daily.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly during acclimation and daily during dosing. Pup weights were recorded after all pups in a litter were delivered and groomed by the dam and again on DL 5.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Following sacrifice
- Anaesthetic used for blood collection: No
- Animals fasted: Yes
- How many animals: 5/sex/dose
- Parameters checked: erythorocycte count, hematocrit, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean corpuscular volume, total leukocyte count, differential leukocyte count, platelet count, mean platelet volume, cell morphology.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Following sacrifice
- Animals fasted: Yes
- How many animals: 5/sex/dose
- Parameters checked: total protein, triglycerides, albumin, globulin, albumin/globulin ratio, glucose, cholesterol, total bilirubin, urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, calcium, phosphorus, sodium, potassium, chloride.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: On one occasion during the study, shortly before scheduled sacrifice.
- Dose groups that were examined: A functional observational battery was conducted on five male and five female rats per group.
- Battery of functions tested: The following parameters were assessed in the functional observational battery: lacrimation, salivation, palpebral closure, prominence of the eye, pupillary reaction to light, piloerection, respiration, urination, defecation, sensorimotor responses to visual, auditory tactile and paulful stimuli, reactions to handeling behavior in the open field, gait pattern in the open field, severity of gait abnormalities, air righting reaction, visual placing response and landing foot splay, forelimb and hindlimb grip strength. Each pup litter was evaluated for viability at least twice daily.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No, females were subjected to necropsy after birthing pups.
- Number of corpora lutea: No
- Number of implantations: No
- Number of early resorptions: No
- Number of late resorptions: No
- Other:

Fetal examinations:

Pups were sacrificed on Day 5 of lactation.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: body weight (DL 1 and 5), clinical signs (once daily), gross necropsy, # liveborn, # stillborn, viability index

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead

Indices:

The following indices were calculated: Number of dams delivering litters, duration of gestation, averages for implantation sites per delivered litter, gestation index, numbers of dams with stillborn pups, and dams with all pups dying.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
All estrou, mating and fertility parameters were unaffected at all dose levels. Absolute liver weights were significantly increased in 250 mg/kg/day-treated females. The ratio of liver weight to brain weight was significantly increased in the 250 mg/kg/day-treated females. Increased kidney weights were noted in the 250 mg/kg group females. Minimal or mild hypertrophy of centrilobular hepatocytes was also observed in most females treated at 250 mg/kg/day.The hypertrophy was due to an increased amount of finely granular, dense eosinophilic cytoplasm. Microscopic examination of the thymus revealed an increased incidence and severity of atrophy of the thymic lobules in female rats treated at 250 mg/kg/day. No microscopic changes were noted in the kidneys upon histopathology.

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 50 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:not examined

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

50 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reduction in number of live offspring

fetal/pup body weight changes

Overall developmental toxicity

Key result

Developmental effects observed:

yes

Lowest effective dose / conc.:

250 mg/kg bw/day (nominal)

Treatment related:

yes

Relation to maternal toxicity:

developmental effects as a secondary non-specific consequence of maternal toxicity effects

Dose response relationship:

yes

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the developmental NOAEL is 50 mg/kg/day.

Executive summary:

The subacute oral toxicity and potential for reproductive and developmental toxicity (screening level) of T-7599 (white powder, purity >99.9%, CASRN 34454-97-2, Lot 6) was evaluated in Sprague-Dawley rats following repeated oral doses. This study was performed in compliance with OECD GLP (1998), US FDA GLP 21 CFR 58, and Japanese MHW GLP (1997). The study method was based on OECD 422 (1996) and US EPA In Vivo Reproductive and Mutagenicity Tests (1982). T-7599 was prepared in 0.5% carboxymethylcellulose (CMC) for the first 4 days of the study and then was prepared in 1.0% CMC for the remainder of the study. Rats (15/sex/dose) received repeated doses of 0 (vehicle), 10, 50, or 250 mg/kg/day via oral gavage at a dose volume of 10 mL/kg. Male rats received their respective doses once daily beginning at 14 days before cohabitation and continuing after the cohabitation period for minimum total of 28 days. Female rats received their respective doses once daily beginning at 14 days before cohabitation and continuing until Day 5 of lactation (LD 5). Within each dosage group, one male rat was assigned per one female rat during the cohabitation period (maximum of 14 days). Mating was confirmed by examining female rats for spermatozoa in vaginal smears and/or absence of copulatory plug. After 36 days of dosage, all males were euthanized. On LD 5 and 6, all surviving pups and females were euthanized, respectively. Parameters evaluated: clinical observations (at least daily), body weight (daily during dosing), food consumption (at least weekly), estrous cycling, litter sizes, pup viability, clinical and necropsy observations of pups, maternal behavior, functional observational battery (5/sex/dose), and hematology and clinical biochemistry (5/sex/dose). Gross necropsy and histological evaluation of select organs were performed on males and females. All males and females survived. Significant increases in excess salivation, perioral substance and urine-stained abdominal fur was observed in the 250 mg/kg/day treated male rats. No abnormal clinical observations were noted in female rats. Body weight gains and absolute and relative food consumption were significantly reduced in the 50 and 250 mg/kg/day-treated males. Body weight gains of the females were significantly reduced during the precohabitation period in the 250 mg/kg/day dose group, but body weights and body weight gains were not significantly affected during gestation or lactation. Terminal body weights of the female rats were reduced in the 250 mg/kg/day dose group. No gross abnormalities were observed upon necropsy. Absolute liver weights were significantly increased in 250 mg/kg/day-treated males and females. Liver weights relative to terminal body weights were significantly increased in the 50 and 250 mg/kg/day treated males. The ratio of liver weight to brain weight was significantly increased in the 250 mg/kg/day-treated males and females. Microscopic examination revealed minimal or mild enlargement (hypertrophy) of centrilobular hepatocytes in most males treated at 250 mg/kg/day and in 4/10 males treated at 50 mg/kg/day. Also in three 250 mg/kg/day-treated males, necrosis of individual enlarged hepatocytes was observed in the centrilobular areas. Minimal or mild hypertrophy of centrilobular hepatocytes was also observed in most females treated at 250 mg/kg/day. In both males and females, the hypertrophy was due to an increased amount of finely granular, dense eosinophilic cytoplasm. In both males and females, increased kidney weights were noted in the 250 mg/kg group and in males alone in the 50 mg/kg group. No microscopic changes were noted in the kidneys (male or female) upon histopathology. Microscopic examination of the thymus revealed an increased incidence and severity of atrophy of the thymic lobules in female rats treated at 250 mg/kg/day. No toxicologically relevant findings in the organ weight and histopathology of the thymus in male rats. Microscopic examination of the stomach revealed focal erosions in the pyloric glandular mucosa of 2 males in the 250 mg/kg/day group. Minimal to mild edema/inflammation of the submucosa of the nonglandular and glandular areas was observed in 2 control group males, 2 males in the 50 mg/kg/day group, and 1 male in the 250 mg/kg/day group. Moderate edema/inflammation of the submucosa of the nonglandular and glandular areas was observed in 2 males in the 250 mg/kg/day group. Thus, the edema/inflammation was observed at a slightly higher incidence and severity in the 250 mg/kg/day dose group. No microscopic changes in the gastrointestinal tract of females were noted. There were no abnormal clinical or necropsy observations in the F1 generation pups. The number of liveborn pups was significantly reduced and the number of stillborn pups was significantly increased in the 250 mg/kg dosage group. The viability index and number of pups surviving per liter on postpartum day 5 were significantly reduced in the 250 mg/kg dosage group. Pup body weight was also reduced in the 250 mg/kg dosage group on postpartum days 1 and 5. Based on the results of the study, the repeat dose toxicity No Observed Adverse Effect Level (NOAEL) for the test article is 50 mg/kg/day. The male and female reproductive NOAEL is 250 mg/kg/day and the developmental NOAEL is 50 mg/kg/day.

The test article is classified as STOT RE 2 (for liver effects) and Repr. 2 according to CLP criteria. The thymus effects seen in the longer-dosed females do not lead to classification as the LOAEL is outside the guidance values for STOT RE 2. The dose at which the liver and thymus effects occurred is outside the guidance values for R48/22.